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dc.contributor.authorTullius, Tomen_US
dc.contributor.authorIngle, Shaktien_US
dc.contributor.authorAzad, Roberten_US
dc.date.accessioned2018-08-14T15:07:42Z
dc.date.available2018-08-14T15:07:42Z
dc.identifier.citationTom Tullius, Shakti Ingle, Robert Azad. "Deuterated nucleotides as chemical probes of RNA structure: a detailed protocol for the enzymatic synthesis of a complete set of nucleotides specifically deuterated at ribose carbons." ScienceOpen Research, https://doi.org/10.14293/S2199-1006.1.SOR-LIFE.ALCJCN.v1
dc.identifier.issn2199-1006
dc.identifier.urihttps://hdl.handle.net/2144/30786
dc.description.abstractWe describe here a detailed protocol for the synthesis of ribonucleotides specifically deuterated at each ribose carbon atom. We synthesized 20 specifically deuterated ribonucleotides: ATP, CTP, GTP, and UTP, each of which contained one of five deuterated riboses (either 1′-D, 2″-D, 3′-D, 4′-D, or 5′,5″-D2). Our synthetic approach is inspired by the pioneering work of Tolbert and Williamson, who developed a method for the convenient one-pot enzymatic synthesis of nucleotides (Tolbert, T. J. and Williamson, J. R. (1996) J. Am. Chem. Soc. 118, 7929–7940). Our protocol consists of a comprehensive list of required chemical and enzymatic reagents and equipment, detailed procedures for enzymatic assays and nucleotide synthesis, and chromatographic procedures for purification of deuterated nucleotides. As an example of the utility of specifically deuterated nucleotides, we used them to synthesize specifically deuterated sarcin/ricin loop (SRL) RNA and measured the deuterium kinetic isotope effect on hydroxyl radical cleavage of the SRL.en_US
dc.description.urihttps://www.scienceopen.com/document/read?vid=eb44f1b0-c408-4336-a2c0-aed203250898
dc.relation.ispartofScienceOpen Research
dc.relation.isversionof10.14293/S2199-1006.1.SOR-LIFE.ALCJCN.v1
dc.rightsCopyright statement: © 2015 Azad et al. License: This work has been published open access under Creative Commons Attribution License CC BY 4.0, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Conditions, terms of use and publishing policy can be found at www.scienceopen.com.en_US
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/
dc.subjectBiochemistryen_US
dc.subjectMolecular biologyen_US
dc.subjectBiophysicsen_US
dc.subjectRNAen_US
dc.subjectRibonucleotide synthesisen_US
dc.subjectHydroxyl radical cleavageen_US
dc.subjectSpecifically deuterated nucleotidesen_US
dc.titleDeuterated nucleotides as chemical probes of RNA structure: a detailed protocol for the enzymatic synthesis of a complete set of nucleotides specifically deuterated at ribose carbonsen_US
dc.typeArticleen_US
dc.description.versionPublished versionen_US
pubs.elements-sourcecrossrefen_US
pubs.notesEmbargo: No embargoen_US
pubs.organisational-groupBoston Universityen_US
pubs.organisational-groupBoston University, College of Arts & Sciencesen_US
pubs.organisational-groupBoston University, College of Arts & Sciences, Department of Chemistryen_US
pubs.publication-statusPublished onlineen_US
dc.date.online2015-05-29
dc.date.online2015-05-29
dc.identifier.orcid0000-0003-4425-796X (Tullius, Tom)


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Copyright statement: © 2015 Azad et al.

License: This work has been published open access under Creative Commons Attribution License CC BY 4.0, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Conditions, terms of use and publishing policy can be found at www.scienceopen.com.
Except where otherwise noted, this item's license is described as Copyright statement: © 2015 Azad et al. License: This work has been published open access under Creative Commons Attribution License CC BY 4.0, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Conditions, terms of use and publishing policy can be found at www.scienceopen.com.